Title: Differential requirement of EGF receptor and its tyrosine kinase for AP-1 transactivation induced by EGF and TPA.
Authors: Li, Jingxia; Ma, Cuiling; Huang, Yi; Luo, Jia; Huang, Chuanshu
Published In Oncogene, (2003 Jan 16)
Abstract: The transcription factor activator protein-1 (AP-1) has been implicated in a large variety of biological processes including cell differentiation, proliferation, apoptosis and oncogenic transformation. It is thought that the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced AP-1 activity is because of the activation of the PKC/MAPK/AP-1 pathway, although the detailed molecular mechanism has not been fully characterized. The tyrosine kinases of epidermal growth factor receptor (EGFR) lie at the head of a complex of signal transduction cascade that modulates cell proliferation, survival, adhesion, migration and differentiation. Currently, little is known about whether EGFR or its tyrosine kinase is necessary for TPA-induced AP-1 activation. In the present study, we investigated this issue using a well-characterized mouse fibroblast B82 cell line, which is devoid of the EGFR, and its stable transfectants with either wild-type EGFR (B82L) or tyrosine kinase-deficient EGFR (mutation at Lys-721) (B82M721). We demonstrated that the TPA or epidermal growth factor (EGF) induced AP-1 activation in the B82L cells that express wild-type EGFR, but not in the B82 cell, whereas autophosphorylation at tyrosine(1173) of EGFR in B82L cells was only induced by EGF, but not TPA. The expression of tyrosine kinase-deficient EGFR (mutation at Lys-721) (B82M721) resulted in deficiency of AP-1 induction in cellular response to EGF, while TPA treatment led to fully AP-1 activation. Furthermore, the mutation at Lys-721 of EGFR resulted in impairing of EGFR autophosphorylation at tyrosine(1173) induced by EGF. Based on these results, we conclude that TPA-induced AP-1 activation requires the basal level-EGFR protein, but not EGFR tyrosine kinase and EGFR autophosphorylation at tyrosine(1173), whereas both EGFR tyrosine kinase and EGFR autophosphorylation at Y(1173) play a critical role in EGF-induced AP-1 activation.
PubMed ID: 12527890
MeSH Terms: Animals; Cells, Cultured; Enzyme Activation/drug effects; Epidermal Growth Factor/metabolism; Epidermal Growth Factor/pharmacology*; Fibroblasts; Humans; Lysine/genetics; Mice; Mitogen-Activated Protein Kinases/drug effects; Mitogen-Activated Protein Kinases/metabolism; Phosphorylation; Point Mutation; Receptor, Epidermal Growth Factor/genetics; Receptor, Epidermal Growth Factor/metabolism*; Tetradecanoylphorbol Acetate/pharmacology*; Transcription Factor AP-1/drug effects; Transcription Factor AP-1/genetics*; Transcription Factor AP-1/metabolism; Transcriptional Activation/drug effects*; Transfection; Tyrosine/metabolism